From Cell Line to Fillet: A Review of Biological and Engineering Strategies in Cultivated Seafood

Abstract

Due to the increasing world population, traditional commercial sources of protein will soon be insufficient to meet global nutritional demands. To address this and other anthropological effects, including carbon emissions, land and freshwater usage and overfishing associated with traditional agriculture, aquaculture and wild-catch fisheries, cultivated meats have been proposed as a novel protein source, rather than animal slaughter. Currently, despite significant investment, the cultivated seafood industry is floundering to scale production. This is due to technological failures to adapt cells to suspension for integration into bioreactors, lack of appropriate scaffold materials and scalable techniques to support eventual product formation. To overcome these issues, it is crucial for the industry to choose methodologies and materials that are tailored to species-specific cells to compensate for differences in the sensitivity and behaviour of cells within different bioreactor systems. Moreover, for regulatory approval and consumer acceptance, appropriate materials and edibility are required for selecting scaffolds. Ultimately, the cultivated seafood industry must address the current limitations in production efficiency to become a viable alternative food source. To do so, the industry should focus on using the recent advancements from both industry and academia including methods and technologies in tissue engineering and the adaption of cells to suspension. This review focuses on cultivated seafood as an alternative to aquaculture and wild-catch fisheries and discusses the bottlenecks and potential solutions for the industry to upscale production.

Keywords

bioreactors / cultivated seafood / scaffolds / suspension culture

References

[1] Abdulhussain, R., A. Adebisi, B. R. Conway, and K. Asare-Addo. 2023. “Electrospun Nanofibers: Exploring Process Parameters, Polymer Selection, and Recent Applications in Pharmaceuticals and Drug Delivery.” Journal of Drug Delivery Science and Technology 90: 105156. https://doi.org/10.1016/j.jddst.2023.105156.

[2] Achinas, S., J. Mulder, and G. J. W. Euverink. 2022. “Chapter 27—A Biotechnological Overview of Syngas Fermentation.” In Handbook of Biofuels, edited by S. Sahay, 511–527. Academic Press. https://doi.org/10.1016/B978-0-12-822810-4.00027-0.

[3] Alasibi, S., M. Kazir, Á. Israel, and Y. D. Livney. 2024. “Algal Protein-Based 3D-printed Fish-Analogs as a New Approach for Sustainable Seafood.” Current Research in Food Science 9: 100905. https://doi.org/10.1016/j.crfs.2024.100905.

[4] Alblawi, A., A. S. Ranjani, H. Yasmin, S. Gupta, A. Bit, and M. Rahimi-Gorji. 2020. “Scaffold-Free: A Developing Technique in Field of Tissue Engineering.” Computer Methods and Programs in Biomedicine 185: 105148. https://doi.org/10.1016/j.cmpb.2019.105148.

[5] Al-Mashhadani, M. K. H., S. J. Wilkinson, and W. B. Zimmerman. 2015. “Airlift Bioreactor for Biological Applications With Microbubble Mediated Transport Processes.” Chemical Engineering Science 137: 243–253. https://doi.org/10.1016/j.ces.2015.06.032.

[6] American Type Culture Collection. 2026. RTgill-W1 (CRL-2523). https://www.atcc.org/products/crl-2523.

[7] Appiani, M., C. Cattaneo, and M. Laureati. 2025. “Plant-Based Fish Analogues vs. Fish: Assessment of Consumer Perception, Acceptance, and Attitudes.” Food Quality and Preference 123: 105329. https://doi.org/10.1016/j.foodqual.2024.105329.

[8] Assis, M. d, D. G. N. Nina, A. Souza, et al. (2024). Marine Biomaterials for Tissue Engineering: Developing Scaffolds for Regenerative Medicine. https://chemistryforsustainability.org/publication/marine-biomaterials-tissue-engineering-developing-scaffolds-regenerative-medicine.

[9] Banavar, A., R. Sarkarat, A. Amirvaresi, et al. 2024. “Decellularized Banana Leaves: Eco-Friendly Scaffolds for Cell-Based Seafood.” Frontiers in Sustainable Food Systems 8: 1–13. https://doi.org/10.3389/fsufs.2024.1341151.

[10] Batish, I.2022. “Developing Serum-Free Media via Bioprocessing Novel Materials for Cultivated Seafood Products [Virginia Tech].” Hampton. https://vtechworks.lib.vt.edu/items/62cb04d7-e822-4d76-bf85-d9f1f4eb71a3.

[11] Bear-McGuinness, L.2026. “Wildtype's Cultivated Salmon Becomes First of Its Kind to Receive FDA Approval.” https://www.technologynetworks.com/applied-sciences/online-events/advances-in-food-beverage-testing-2026-407449.

[12] Bellani, C. F., J. Ajeian, L. Duffy, M. Miotto, L. Groenewegen, and C. J. Connon. 2020. “Scale-Up Technologies for the Manufacture of Adherent Cells.” Frontiers in Nutrition 7: 1–14. https://doi.org/10.3389/fnut.2020.575146.

[13] Benito-Jardón, M., N. Strohmeyer, S. Ortega-Sanchís, et al. 2020. “αv-Class Integrin Binding to Fibronectin Is Solely Mediated by RGD and Unaffected by an RGE Mutation.” Journal of Cell Biology 219, no. 12: 1–17. https://doi.org/10.1083/jcb.202004198.

[14] Bilodeau, A. R.2024. “The Pros and Cons of Adherent Versus Suspension Cell Culture.” BioPharm International 37, no. 1: 18–22.

[15] Bodiou, V., P. Moutsatsou, and M. J. Post. 2020. “Microcarriers for Upscaling Cultured Meat Production.” Frontiers in Nutrition 7: 1–16. https://doi.org/10.3389/fnut.2020.00010.

[16] Boppart, M. D., and Z. S. Mahmassani. 2019. “Integrin Signaling: Linking Mechanical Stimulation to Skeletal Muscle Hypertrophy.” American Journal of Physiology-Cell Physiology 317, no. 4: C629–C641. https://doi.org/10.1152/ajpcell.00009.2019.

[17] Caliari, S. R., and J. A. Burdick. 2016. “A Practical Guide to Hydrogels for Cell Culture.” Nature Methods 13, no. 5: 405–414. https://doi.org/10.1038/nmeth.3839.

[18] CellBase. 2026. “MOR Cell.bio Marbled Goby MUSC.” https://cellbase.com/collections/seafood-cell-lines/products/mor-cell-biomarbled-goby-musc-mgm_c1.

[19] Chaudhry, M. A.2024. “Lessons in Bioreactor Scale-Up: Part 1—Exploring Introductory Principles.” https://www.bioprocessintl.com/bioreactors/lessons-in-bioreactor-scale-up-part-1-mdash-exploring-introductory-principles.

[20] Chen, G., and N. Kawazoe. 2023. “Polymeric and Biomimetic ECM Scaffolds for Tissue Engineering Applications.” In Polymeric Biomaterials for Tissue Regeneration: From Surface/Interface Design to 3D Constructs, edited by C. Gao, 41–60. Springer Nature Singapore. https://doi.org/10.1007/978-981-99-6948-7_2.

[21] Chen, X. Y., J. Y. Chen, X. M. Tong, J. G. Mei, Y. F. Chen, and X. Z. Mou. 2020. “Recent Advances in the Use of Microcarriers for Cell Cultures and Their Ex Vivo and In Vivo Applications.” Biotechnology Letters 42, no. 1: 1–10. https://doi.org/10.1007/s10529-019-02738-7.

[22] Ciriza, J., A. Rodríguez-Romano, I. Nogueroles, et al. 2021. “Borax-Loaded Injectable Alginate Hydrogels Promote Muscle Regeneration In Vivo After an Injury.” Materials Science and Engineering: C 123: 112003. https://doi.org/10.1016/j.msec.

[23] Coleman, B., C. Van Poucke, B. Dewitte, et al. 2022. “Potential of Microalgae as Flavoring Agents for Plant-Based Seafood Alternatives.” Future Foods 5: 100139. https://doi.org/10.1016/j.fufo.2022.100139.

[24] Collins, M. N., G. Ren, K. Young, S. Pina, R. L. Reis, and J. M. Oliveira. 2021. “Scaffold Fabrication Technologies and Structure/Function Properties in Bone Tissue Engineering.” Advanced Functional Materials 31, no. 21: 2010609. https://doi.org/10.1002/adfm.202010609.

[25] Csapo, R., M. Gumpenberger, and B. Wessner. 2020. “Skeletal Muscle Extracellular Matrix – What Do We Know About Its Composition, Regulation, and Physiological Roles? A Narrative Review.” Frontiers in Physiology 11: 2020. https://doi.org/10.3389/fphys.2020.00253.

[26] Daly, A. C., M. E. Prendergast, A. J. Hughes, and J. A. Burdick. 2021. “Bioprinting for the Biologist.” Cell 184, no. 1: 18–32. https://doi.org/10.1016/j.cell.2020.12.002.

[27] Derakhti, S., S. H. Safiabadi-Tali, G. Amoabediny, and M. Sheikhpour. 2019. “Attachment and Detachment Strategies in Microcarrier-Based Cell Culture Technology: A Comprehensive Review.” Materials Science and Engineering: C 103: 109782. https://doi.org/10.1016/j.msec.

[28] Drochon, A., R. Lesieur, and M. Durand. 2022. “Fluid Dynamics Characterisation of a Rotating Bioreactor for Tissue Engineering.” Medical Engineering & Physics 105: 103831. https://doi.org/10.1016/j.medengphy.2022.103831.

[29] Engler, A. J., H. L. Sweeney, D. E. Discher, and J. E. Schwarzbauer. 2007. “Extracellular Matrix Elasticity Directs Stem Cell Differentiation.” Journal of Musculoskeletal & Neuronal Interactions 7, no. 4: 335.

[30] Escamilla Silva, E. M., and A. M. Mendoza-Martínez. 2013. “Airlift Bioreactors: Hydrodynamics and Rheology Application to Secondary Metabolites Production.” In Mass Transfer—Advances in Sustainable Energy and Environment Oriented Numerical Modeling, edited by H. Nakajima. IntechOpen. https://doi.org/10.5772/53711.

[31] faCellitate. 2022. “In Vitro Cell Culture Techniques: Adherent Culture vs. Suspension Culture.” https://facellitate.com/in-vitro-cell-culture-techniques-adherent-culture-vs-suspension-culture/.

[32] Fasciano, S., A. Wheba, C. Ddamulira, and S. Wang. 2024. “Recent Advances in Scaffolding Biomaterials for Cultivated Meat.” Biomaterials Advances 162: 213897. https://doi.org/10.1016/j.bioadv.2024.213897.

[33] Feng, S., S. Dai, Z. Wei, J. Wang, N. Xiang, and P. Shao. 2024. “Soy Conglycinin Amyloid Fibril and Chitosan Complex Scaffold for Cultivated Meat Application.” Food Hydrocolloids 153: 110017. https://doi.org/10.1016/j.foodhyd.2024.110017.

[34] Fonkwe, L. G., G. Narsimhan, and A. S. Cha. 2003. “Characterization of Gelation Time and Texture of Gelatin and Gelatin–Polysaccharide Mixed Gels.” Food Hydrocolloids 17, no. 6: 871–883. https://doi.org/10.1016/S0268-005X(03)00108-5.

[35] Food Standards Australia New Zealand. 2026. “A1269—Cultured Quail as a Novel Food.” https://www.foodstandards.gov.au/food-standards-code/applications/A1269-Cultured-Quail-as-a-Novel-Food.

[36] García de la serrana, D., M. Codina, E. Capilla, et al. 2014. “Characterisation and Expression of Myogenesis Regulatory Factors During in Vitro Myoblast Development and In Vivo Fasting in the Gilthead Sea Bream (Sparus Aurata).” Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 167: 90–99. https://doi.org/10.1016/j.cbpa.2013.10.020.

[37] García-Pérez, I., I. Rodríguez, A. Rubio, et al. 2025. “Characterization of Myogenesis in European Sea Bass (Dicentrarchus Labrax) Using Primary White Muscle Cell Cultures.” Scientific Reports 15, no. 1: 45778. https://doi.org/10.1038/s41598-025-28654-7.

[38] Ghafouri Azar, M., L. Wiesnerova, J. Dvorakova, et al. 2023. “Optimizing PCL/PLGA Scaffold Biocompatibility Using Gelatin From Bovine, Porcine, and Fish Origin.” Gels 9, no. 11: 900. https://doi.org/10.3390/gels9110900.

[39] Gomez Romero, S., and N. Boyle. 2023. “Systems Biology and Metabolic Modeling for Cultivated Meat: A Promising Approach for Cell Culture Media Optimization and Cost Reduction.” Comprehensive Reviews in Food Science and Food Safety 22, no. 4: 3422–3443. https://doi.org/10.1111/1541-4337.13193.

[40] Good Food Institute. 2026. “Ensuring a Clear Path to Market.” https://gfi.org/ensuring-a-clear-path-to-market/.

[41] Goswami, M., R. Ovissipour, C. Bomkamp, et al. 2024. “Cell-Cultivated Aquatic Food Products: Emerging Production Systems for Seafood.” Journal of Biological Engineering 18, no. 1: 43. https://doi.org/10.1186/s13036-024-00436-1.

[42] Grounds, M. D., L. Sorokin, and J. White. 2005. “Strength at the Extracellular Matrix–Muscle Interface.” Scandinavian Journal of Medicine & Science in Sports 15, no. 6: 381–391. https://doi.org/10.1111/j.1600-0838.2005.00467.x.

[43] Guzman-Puyol, S., J. A. Heredia-Guerrero, L. Ceseracciu, et al. 2016. “Low-Cost and Effective Fabrication of Biocompatible Nanofibers From Silk and Cellulose-Rich Materials.” ACS Biomaterials Science & Engineering 2, no. 4: 526–534. https://doi.org/10.1021/acsbiomaterials.5b00500.

[44] Hartman, M.2025. Re: Cell Culture Consultation CCC 000005. U.S Food & Drug Administration.

[45] Hu, H., L. Krishaa, and E. L. S. Fong. 2023. “Magnetic Force-Based Cell Manipulation for In Vitro Tissue Engineering.” APL Bioengineering 7, no. 3: 031504. https://doi.org/10.1063/5.0138732.

[46] Huang, H., J. N. Bogstie, and H. J. Vogel. 2012. “Biophysical and Structural Studies of the Human Calcium- and Integrin-Binding Protein Family: Understanding Their Functional Similarities and Differences.” Biochemistry and Cell Biology 90, no. 5: 646–656. https://doi.org/10.1139/o2012-021%25M.

[47] Hwang, H., and S. J. Nam. 2021. “The Influence of Consumers’ Knowledge on Their Responses to Genetically Modified Foods.” GM Crops & Food 12, no. 1: 146–157. https://doi.org/10.1080/21645698.2020.1840911.

[48] Iyer, S. R., N. Udpa, and Y. Gao. 2015. “Chitosan Selectively Promotes Adhesion of Myoblasts over Fibroblasts.” Journal of biomedical materials research. Part A 103, no. 6: 1899–1906. https://doi.org/10.1002/jbm.a.35075.

[49] Jana, S., P. Das, P. R. Ghosh, and S. K. Nandi. 2024. “Collagen and Gelatin From Fish Processing By-Products for Biomedical Applications.” In Fish Waste to Valuable Products, edited by S. Maqsood, M. N. Naseer, S. Benjakul, and A. A. Zaidi, 91–117. Springer Nature Singapore. https://doi.org/10.1007/978-981-99-8593-7_6.

[50] de Jesus, S. S., J. Moreira Neto, and R. Maciel Filho. 2017. “Hydrodynamics and Mass Transfer in Bubble Column, Conventional Airlift, Stirred Airlift and Stirred Tank Bioreactors, Using Viscous Fluid: A Comparative Study.” Biochemical Engineering Journal 118: 70–81. https://doi.org/10.1016/j.bej.2016.11.019.

[51] Jin, X., J. Y. Park, J. S. Lee, U. W. Jung, S. H. Choi, and J. K. Cha. 2023. “Tissue Integration Patterns of Non-Crosslinked and Crosslinked Collagen Membranes: An Experimental In Vivo Study.” Journal of Periodontal & Implant Science 53, no. 3: 207–217. https://doi.org/10.5051/jpis.2203260163.

[52] John, J.2019. BlueNalu Makes a Splash With Groundbreaking Cultivated Yellowtail. Good Food Institute. https://gfi.org/blog/bluenalu-cultivated-yellowtail/.

[53] Johnson, H., and A. Monaco. 2025. “Global Developments in the Regulation of Cultivated Meat: A Comparative Study of the EU, Singapore, US and Australia and New Zealand.” Review of European, Comparative & International Environmental Law 34, no. 2: 496–511. https://doi.org/10.1111/reel.70007.

[54] Joyce, C. M., E. B. Gordon, A. McGivney, et al. 2024. “Methods to Screen the Adhesion of Fish Cells on Plant-, Algal- and Fungal-Derived Biomaterials.” ACS Applied Materials & Interfaces 16, no. 30: 39969–39980. https://doi.org/10.1021/acsami.4c06543.

[55] Jozef, B., Z. R. Zhang, H.-M. Kaltenbach, and K. Schirmer. 2025. “A Systematic Approach Towards Long-Term, Serum-Free Cultivation of Fish Cells With the RTgill-W1 Cell Line as Example.” NAM Journal 1: 100008. https://doi.org/10.1016/j.namjnl.2025.100008.

[56] Jukić, S., D. Bubenik, N. Pavlović, A. J. Tušek, and V. G. Srček. 2016. “Adaptation of CHO Cells in Serum-Free Conditions for Erythropoietin Production: Application of EVOP Technique for Process Optimization.” Biotechnology and Applied Biochemistry 63, no. 5: 633–641. https://doi.org/10.1002/bab.1468.

[57] Kang, H. S., S. Bang, H. Lee, et al. 2025. “Development of Cultivated Fish Meat: Advances in Cellular Agriculture, Biomaterials, and Scaffolding Techniques.” Trends in Food Science & Technology 162: 105088. https://doi.org/10.1016/j.tifs.2025.105088.

[58] Khan, F. A.2022. “Stem Cell Bioreactors: Design, Structure, and Operation of Stem Cell Bioreactors.” In Stem Cell Production: Processes, Practices and Regulations, edited by F. A. Khan, 113–123. Springer Singapore. https://doi.org/10.1007/978-981-16-7589-8_5.

[59] Khan, S., M. Zulfiqar, and S. Imtiaz. 2024. “The Scaling-Up of Lab-Grown Meat Production: A Critical Analysis of Nutritional Profiling, Challenges, and Ethical Implications.” Pakistan Journal of Biochemistry and Molecular Biology 56, no. 2: 2.

[60] Kim, W.-J., Y. Kim, R. Ovissipour, and N. Nitin. 2024. “Plant-Based Biomaterials as Scaffolds for Cellular Agriculture.” Future Foods 10: 100468. https://doi.org/10.1016/j.fufo.2024.100468.

[61] Kraemer, P. M., F. A. Ray, A. R. Brothman, M. F. Barthoidi, and L. S. Cram. 1986. “Spontaneous Immortalization Rate of Cultured Chinese Hamster Cells.” JNCI: Journal of the National Cancer Institute 76, no. 4: 703–709. https://doi.org/10.1093/jnci/76.4.703.

[62] Kulus, M., M. Jankowski, W. Kranc, et al. 2023. “Bioreactors, Scaffolds and Microcarriers and In Vitro Meat Production—Current Obstacles and Potential Solutions.” Frontiers in Nutrition 10: 1–11. https://doi.org/10.3389/fnut.2023.1225233.

[63] Kuppusamy, P., D. Kim, I. Soundharrajan, I. Hwang, and K. C. Choi. 2020. “Adipose and Muscle Cell Co-Culture System: A Novel In Vitro Tool to Mimic the In Vivo Cellular Environment.” Biology 10, no. 1: 1–12. https://doi.org/10.3390/biology10010006.

[64] Lambert, E. G., C. J. O'Keeffe, A. O. Ward, T. A. Anderson, Q. Yip, and P. L. H. Newman. 2024. “Enhancing the Palatability of Cultivated Meat.” Trends in Biotechnology 42, no. 9: 1112–1127. https://doi.org/10.1016/j.tibtech.2024.02.014.

[65] de Lamotte, A., A. Delafosse, S. Calvo, F. Delvigne, and D. Toye. 2017. “Investigating the Effects of Hydrodynamics and Mixing on Mass Transfer Through the Free-Surface in Stirred Tank Bioreactors.” Chemical Engineering Science 172: 125–142. https://doi.org/10.1016/j.ces.2017.06.028.

[66] Lee, D. Y., S. Y. Lee, S. H. Yun, et al. 2022. “Review of the Current Research on Fetal Bovine Serum and the Development of Cultured Meat.” Food Science of Animal Resources 42, no. 5: 775–799. https://doi.org/10.5851/kosfa.2022.e46.

[67] Lee, D. Y., S. H. Yun, J. Lee, et al. 2024. “The Roles of Media Ingredients in Muscle Cell Culture for Cultured Meat Production—A Mini-Review.” Future Foods 10: 100403. https://doi.org/10.1016/j.fufo.2024.100403.

[68] Lee, K.-B., and L. Yang. 2019. “Scaffold-Free Biological Assemblies for Tissue Modeling and Regeneration (United States of America Patent No).” https://techfinder.rutgers.edu/tech/Scaffold-Free_Biological_Assemblies_for_Tissue_Modeling_and_Regeneration.

[69] Lee, Y.-E., E. S. Jeong, Y.-M. Kim, and S. P. Gong. 2024. “Investigation of Scaffold Manufacturing Conditions for 3-dimensional Culture of Myogenic Cell Line Derived From Black Sea Bream (Acanthopagrus Schlegelii).” Cytotechnology 77, no. 1: 18. https://doi.org/10.1007/s10616-024-00676-5.

[70] Li, X., B. Liu, B. Pei, et al. 2020. “Inkjet Bioprinting of Biomaterials.” Chemical Reviews 120, no. 19: 10793–10833. https://doi.org/10.1021/acs.chemrev.0c00008.

[71] Li, Y., T. Xue, S. Wang, et al. 2025. “Establishment of Two Continuous Adult Stem Cell Lines From Muscle and Adipose Tissues of Rainbow Trout (Oncorhynchus Mykiss) for Marbled Fish Meat Production.” Aquaculture Reports 43: 102913. https://doi.org/10.1016/j.aqrep.2025.102913.

[72] Lim, K.-T., D. K. Patel, H. Seonwoo, J. Kim, and J. H. Chung. 2019. “A Fully Automated Bioreactor System for Precise Control of Stem Cell Proliferation and Differentiation.” Biochemical Engineering Journal 150: 107258. https://doi.org/10.1016/j.bej.2019.107258.

[73] Listrat, A., B. Lebret, I. Louveau, et al. 2016. “How Muscle Structure and Composition Influence Meat and Flesh Quality.” Scientific World Journal 2016: 3182746. https://doi.org/10.1155/2016/3182746.

[74] Liu, N., X. Zhang, Q. Guo, T. Wu, and Y. Wang. 2022. “3D Bioprinted Scaffolds for Tissue Repair and Regeneration.” Frontiers in Materials 9: 1–17. https://doi.org/10.3389/fmats.2022.925321.

[75] Liu, S., C.-S. Lau, K. Liang, F. Wen, and S. H. Teoh. 2022. “Marine Collagen Scaffolds in Tissue Engineering.” Current Opinion in Biotechnology 74: 92–103. https://doi.org/10.1016/j.copbio.2021.10.011.

[76] Liu, Z., J. Shi, L. Chen, et al. 2024. “3D Printing of Fish-Scale Derived Hydroxyapatite/Chitosan/PCL Scaffold for Bone Tissue Engineering.” International Journal of Biological Macromolecules 274: 133172. https://doi.org/10.1016/j.ijbiomac.2024.133172.

[77] Long, X., W. Chen, G. Liu, W. Hu, and Q. Tan. 2023. “Establishment and Characterization of a Skeletal Myoblast Cell Line of Grass Carp (Ctenopharyngodon Idellus).” Fish Physiology and Biochemistry 49, no. 5: 1043–1061. https://doi.org/10.1007/s10695-023-01246-w.

[78] Lordon, B., T. Campion, L. Gibot, and G. Gallot. 2024. “Impact of Trypsin on Cell Cytoplasm During Detachment of Cells Studied by Terahertz Sensing.” Biophysical Journal 123, no. 16: 2476–2483. https://doi.org/10.1016/j.bpj.2024.06.011.

[79] Maihemuti, A., H. Zhang, X. Lin, et al. 2023. “3D-Printed Fish Gelatin Scaffolds for Cartilage Tissue Engineering.” Bioactive Materials 26: 77–87. https://doi.org/10.1016/j.bioactmat.2023.02.007.

[80] Marques, D. M. C., M. Jabouille, A. Gusmão, M. Leite, P. Sanjuan-Alberte, and F. C. Ferreira. 2025. “Microalgae-Enriched (Bio)Inks for 3D Bioprinting of Cultured Seafood.” NPJ Science of Food 9, no. 1: 23. https://doi.org/10.1038/s41538-025-00386-y.

[81] Marques, D. M. C., B. D. Pereira, B. Malhão, J. C. Silva, P. Sanjuan-Alberte, and F. C. Ferreira. 2025. “Mimicking the Myoseptum in Cultivated Fish by Manufacturing Edible Microalgae-Rich Nanofibers.” NPJ Science of Food 9, no. 1: 145. https://doi.org/10.1038/s41538-025-00508-6.

[82] Mithra, S., S. Abdul Majeed, S. A. Eisa Abdullah, et al. 2026. “Production of Small-Scale Laboratory-Grown Cell-Based Fish Meat From Asian Seabass Muscle and Fin Cell Lines.” In Vitro Cellular & Developmental Biology. Animal 62: 337–344. https://doi.org/10.1007/s11626-025-01040-3.

[83] Mukhopadhyay, D., and R. De. 2022. “Growth Kinetics and Power Laws Indicate Distinct Mechanisms of Cell-Cell Interactions in the Aggregation Process.” Biophysical Journal 121, no. 3: 481–490. https://doi.org/10.1016/j.bpj.2021.12.030.

[84] Negrete B, Jr, K. L. Ackerly, and A. J. Esbaugh. 2024. “Hypoxia-Acclimation Adjusts Skeletal Muscle Anaerobic Metabolism and Burst Swim Performance in a Marine Fish.” Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 297: 111734. https://doi.org/10.1016/j.cbpa.2024.111734.

[85] Ng, W. L., and J. S. Tan. 2025. “Machine Learning in Cultivated Meat: Enhancing Sustainability, Efficiency, Quality, and Scalability Across the Production Pipeline.” Food and Bioprocess Technology 18, no. 7: 5988–6009. https://doi.org/10.1007/s11947-025-03853-2.

[86] Nickel, N., J. Fitschen, I. Haase, et al. 2024. “Novel Sparging Strategies to Enhance Dissolved Carbon Dioxide Stripping in Industrial Scale Stirred Tank Reactors.” Frontiers in Chemical Engineering 6: 1–10. https://doi.org/10.3389/fceng.2024.1470991.

[87] Nikkhah, A., A. Rohani, M. Zarei, et al. 2023. “Toward Sustainable Culture Media: Using Artificial Intelligence to Optimize Reduced-Serum Formulations for Cultivated Meat.” Science of the Total Environment 894: 164988. https://doi.org/10.1016/j.scitotenv.2023.164988.

[88] Nurul Alam, A. M. M., C.-J. Kim, S.-H. Kim, et al. 2024. “Scaffolding Fundamentals and Recent Advances in Sustainable Scaffolding Techniques for Cultured Meat Development.” Food Research International 189: 114549. https://doi.org/10.1016/j.foodres.2024.114549.

[89] O'Brien, F. J.2011. “Biomaterials & Scaffolds for Tissue Engineering.” Materials Today 14, no. 3: 88–95. https://doi.org/10.1016/S1369-7021(11)70058-X.

[90] Ong, K. J., J. Johnston, I. Datar, V. Sewalt, D. Holmes, and J. A. Shatkin. 2021. “Food Safety Considerations and Research Priorities for the Cultured Meat and Seafood Industry.” Comprehensive Reviews in Food Science and Food Safety 20, no. 6: 5421–5448. https://doi.org/10.1111/1541-4337.12853.

[91] Pajčin, I., T. Knežić, I. Savic Azoulay, et al. 2022. “Bioengineering Outlook on Cultivated Meat Production.” Micromachines 13, no. 3: 1–44. https://doi.org/10.3390/mi13030402.

[92] Palladino, F., P. R. F. Marcelino, A. E. Schlogl, et al. 2024. “Bioreactors: Applications and Innovations for a Sustainable and Healthy Future—A Critical Review.” Applied Sciences 14, no. 20: 9346.

[93] Park, S. M., J. H. Ryoo, H. C. Kwon, and S. G. Han. 2025. “Scaffold Biomaterials in the Development of Cultured Meat: A Review.” Food Science of Animal Resources 45, no. 3: 688–710. https://doi.org/10.5851/kosfa.2025.e13.

[94] Raines, E. W., and K. E. Bornfeldt. 2010. “Integrin α7β1 Compels Smooth Muscle Cells to Maintain Their Quiescence.” Circulation Research 106, no. 3: 427–429. https://doi.org/10.1161/CIRCRESAHA.109.214072.

[95] Rasche, U.2024. “Bioreactors and Fermentors—Powerful Tools for Resolving Cultivation Bottlenecks.” Eppendorf 21: 1–12.

[96] Reiss, J., S. Robertson, and M. Suzuki. 2021. “Cell Sources for Cultivated Meat: Applications and Considerations Throughout the Production Workflow.” International Journal of Molecular Sciences 22, no. 14: 7513. https://doi.org/10.3390/ijms22147513.

[97] Riquelme-Guzmán, C., A. J. Stout, D. L. Kaplan, and J. E. Flack. 2024. “Unlocking the Potential of Cultivated Meat Through Cell Line Engineering.” iScience 27, no. 10: 110877. https://doi.org/10.1016/j.isci.2024.110877.

[98] Rossi, G., and G. Messina. 2014. “Comparative Myogenesis in Teleosts and Mammals.” Cellular and Molecular Life Sciences 71, no. 16: 3081–3099. https://doi.org/10.1007/s00018-014-1604-5.

[99] Ryall, J., A. Ward, and S. Topfer. 2025. Suspension Cultures. Australia Patent No. World Intellectual Property Organisation.

[100] Saad, M. K., J. S. K. Yuen, C. M. Joyce, et al. 2023. “Continuous Fish Muscle Cell Line With Capacity for Myogenic and Adipogenic-Like Phenotypes.” Scientific Reports 13, no. 1: 5098. https://doi.org/10.1038/s41598-023-31822-2.

[101] Santos, A. E. A. d, J. L. Guadalupe, J. D. S. Albergaria, et al. 2024. “Random Cellulose Acetate Nanofibers: A Breakthrough for Cultivated Meat Production.” Frontiers in Nutrition 10: 1–17. https://doi.org/10.3389/fnut.2023.1297926.

[102] Seibert, G. A., V. Feddern, A. P. A. Bastos, A. Kumar, and S. Verruck. 2025. “Trends in Non-Animal Scaffolds for Cultured Meat Structuration.” NPJ Science of Food 9, no. 1: 208. https://doi.org/10.1038/s41538-025-00429-4.

[103] Sen, P., A. Nath, and C. Bhattacharjee. 2017. “9 - Packed-Bed Bioreactor and Its Application in Dairy, Food, and Beverage Industry.” In Current Developments in Biotechnology and Bioengineering, edited by C. Larroche, M. Á. Sanromán, G. Du, and A. Pandey, 235–277. Elsevier. https://doi.org/10.1016/B978-0-444-63663-8.00009-4.

[104] Seo, J. W., W. K. Jung, Y. H. Park, and H. Bae. 2023. “Development of Cultivable Alginate Fibers for An Ideal Cell-Cultivated Meat Scaffold and Production of Hybrid Cultured Meat.” Carbohydrate Polymers 321: 121287. https://doi.org/10.1016/j.carbpol.2023.121287.

[105] Sheridan, W. S., G. P. Duffy, and B. P. Murphy. 2013. “Optimum Parameters for Freeze-Drying Decellularized Arterial Scaffolds.” Tissue Engineering Part C: Methods 19, no. 12: 981–990. https://doi.org/10.1089/ten.TEC.2012.0741.

[106] Shi, H., J. Li, E. Xu, H. Yang, D. Liu, and J. Yin. 2023. “Microscale 3D Printing of Fish Analogues Using Soy Protein Food Ink.” Journal of Food Engineering 347: 111436. https://doi.org/10.1016/j.jfoodeng.2023.111436.

[107] Skrivergaard, S., J. F. Young, N. Sahebekhtiari, et al. 2023. “A Simple and Robust Serum-Free Media for the Proliferation of Muscle Cells.” Food Research International 172: 113194. https://doi.org/10.1016/j.foodres.2023.113194.

[108] Smith, J. A., and E. Mele. 2021. “Electrospinning and Additive Manufacturing: Adding Three-Dimensionality to Electrospun Scaffolds for Tissue Engineering.” Frontiers in Bioengineering and Biotechnology 9: 1–7. https://doi.org/10.3389/fbioe.2021.674738.

[109] Soleymani, S., S. M. Naghib, and M. R. Mozafari. 2024. “An Overview of Cultured Meat and Stem Cell Bioprinting: How to Make It, Challenges and Prospects, Environmental Effects, Society's Culture and the Influence of Religions.” Journal of Agriculture and Food Research 18: 101307. https://doi.org/10.1016/j.jafr.2024.101307.

[110] Stout, A. J., A. B. Mirliani, M. L. Rittenberg, et al. 2022. “Simple and Effective Serum-Free Medium for Sustained Expansion of Bovine Satellite Cells for Cell Cultured Meat.” Communications Biology 5, no. 1: 466. https://doi.org/10.1038/s42003-022-03423-8.

[111] Stout, A. J., M. L. Rittenberg, M. Shub, et al. 2023. “A Beefy-R Culture Medium: Replacing Albumin With Rapeseed Protein Isolates.” Biomaterials 296: 122092. https://doi.org/10.1016/j.biomaterials.2023.122092.

[112] Tan, J. S., and W. L. Ng. 2025. “3D Printing of Salmon Sashimi Analogues Using Fish-Derived Food Inks: Printability, Texture, and Nutritional Evaluation.” Food and Bioprocess Technology 18, no. 10: 8539–8553. https://doi.org/10.1007/s11947-025-03943-1.

[113] Tanaka, R., K. Sakaguchi, A. Yoshida, H. Takahashi, Y. Haraguchi, and T. Shimizu. 2022. “Production of Scaffold-Free Cell-Based Meat Using Cell Sheet Technology.” NPJ Science of Food 6, no. 1: 41. https://doi.org/10.1038/s41538-022-00155-1.

[114] Tay, J. U., C. Zhou, H. W. Lee, Y. Lu, and D. Huang. 2023. “3D Printing of Salmon Fillet Mimic: Imparting Printability via High-Pressure Homogenization and Post-Printing Texturization via Transglutaminase.” Food Hydrocolloids 140: 108564. https://doi.org/10.1016/j.foodhyd.2023.108564.

[115] Taylor, L., M. Wankell, P. Saxena, C. McFarlane, and L. Hebbard. 2022. “Cell Adhesion an Important Determinant of Myogenesis and Satellite Cell Activity.” Biochimica et Biophysica Acta (BBA)—Molecular Cell Research 1869, no. 2: 119170. https://doi.org/10.1016/j.bbamcr.2021.119170.

[116] Thummarati, P., W. Laiwattanapaisal, R. Nitta, M. Fukuda, A. Hassametto, and M. Kino-Oka. 2023. “Recent Advances in Cell Sheet Engineering: From Fabrication to Clinical Translation.” Bioengineering (Basel, Switzerland) 10, no. 2: 1–24. https://doi.org/10.3390/bioengineering10020211.

[117] Trace, A., M. Wankell, C. McFarlane, and L. Hebbard. 2025. “The Challenges of Using Fish Cells for Cultivated Seafood Production.” Food Science and Biotechnology 34: 1565–1579. https://doi.org/10.1007/s10068-024-01786-8.

[118] Traylor, S. D., E. F. Granek, M. Duncan, and S. M. Brander. 2024. “From the Ocean to Our Kitchen Table: Anthropogenic Particles in the Edible Tissue of U.S. West Coast Seafood Species.” Frontiers in Toxicology 6: 1–13. https://doi.org/10.3389/ftox.2024.1469995.

[119] Urtti, A.2024. “Bioreactors: A Comprehensive Overview.” Pharmaceutical Bioprocessing 12, no. 6: 246–247. https://doi.org/10.37532/2048-9145.2024.12(6).

[120] Vendruscolo, F., M. J. Rossi, W. Schmidell, and J. L. Ninow. 2012. “Determination of Oxygen Solubility in Liquid Media.” International Scholarly Research Notices 2012, no. 1: 601458. https://doi.org/10.5402/2012/601458.

[121] Vivek, V., F. N. Eka, and W. Chew. 2022. “Mixing Studies in an Unbaffled Bioreactor Using a Computational Model Corroborated With In-Situ Raman and Imaging Analyses.” Chemical Engineering Journal Advances 9: 100232. https://doi.org/10.1016/j.ceja.2021.100232.

[122] Waler, T.2024. “Bioreactors: The Heart of Pharmacautical Bioprocessing.” Perspective - Pharmceutical Bioprocessing 12, no. 4: 208–209.

[123] Wang, F., X. Cai, Y. Shen, and L. Meng. 2023. “Cell-Scaffold Interactions in Tissue Engineering for Oral and Craniofacial Reconstruction.” Bioactive Materials 23: 16–44. https://doi.org/10.1016/j.bioactmat.2022.10.029.

[124] Watson, E.2023. Brief: Cultivated Seafood Startup BlueNalu Bags $33.5m in Series B Round. https://agfundernews.com/cultivated-seafood-startup-bluenalu-bags-33-5m-in-series-b-round.

[125] Watson, E.2025. FDA Clears Wildtype's Cell-Cultivated Salmon for US debut. https://agfundernews.com/fda-clears-wildtypes-cell-cultivated-salmon-for-us-debut.

[126] Wei, Y., D. Song, J. Teng, et al. 2025. “Establishment of a Continuous Atlantic Salmon Muscle Cell Line for Cell-Cultured Fish Meat and Characterization of Its Adipogenic Transdifferentiation Induced by n-3 and n-6 Fatty Acids.” Food Research International 221: 117610. https://doi.org/10.1016/j.foodres.2025.117610.

[127] World Intellectual Property Organisation. 2022. Shiok Meat Pioneer in Cellular Agriculture. https://www.wipo.int/en/web/ip-advantage/w/stories/shiok-meat-pioneer-in-cellular-agriculture.

[128] Xia, P., H. Miyajima, and S. Fujita. 2025. “Development of Biomimetic Edible Scaffolds for Cultured Meat Based on the Traditional Freeze-Drying Method for Ito-Kanten (Japanese Freeze-Dried Agar).” Gels 11, no. 4: 299. https://doi.org/10.3390/gels11040299.

[129] Xue, T., H. Zheng, Y. Zhao, et al. 2024. “A Spontaneously Immortalized Muscle Stem Cell Line (EfMS) From Brown-Marbled Grouper for Cell-Cultured Fish Meat Production.” Communications Biology 7, no. 1: 1697. https://doi.org/10.1038/s42003-024-07400-1.

[130] Yang, O., S. Prabhu, and M. Ierapetritou. 2019. “Comparison Between Batch and Continuous Monoclonal Antibody Production and Economic Analysis.” Industrial & Engineering Chemistry Research 58, no. 15: 5851–5863. https://doi.org/10.1021/acs.iecr.8b04717.

[131] Yasu, R.2024. “Upstream and Downstream Processing in Pharmaceutical Bioprocessing.” Pharmaceutical Bioprocessing 12, no. 4: 210–211. https://doi.org/10.37532/2048-9145.2024.12(4).210-211.

[132] Zheng, Z., B. Chen, X. Liu, et al. 2024. “Novel Eel Skin Fibroblast Cell Line: Bridging Adherent and Suspension Growth for Aquatic Applications Including Virus Susceptibility.” Biology 13, no. 12: 1068.

[133] Zhou, H., L. S. W. Loo, F. Y. T. Ong, et al. 2025. “Cost-Effective Production of Meaty Aroma From Porcine Cells for Hybrid Cultivated Meat.” Food Chemistry 473: 142946. https://doi.org/10.1016/j.foodchem.2025.142946.

[134] Zhu, K., H. Wang, H. Wang, et al. 2014. “Characterization of Muscle Morphology and Satellite Cells, and Expression of Muscle-Related Genes in Skeletal Muscle of Juvenile and Adult Megalobrama Amblycephala.” Micron 64: 66–75. https://doi.org/10.1016/j.micron.2014.03.009.

Rights & permissions

2026 The Author(s). Food Bioengineering published by John Wiley & Sons Australia, Ltd. on behalf of State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology.